2,4,6-trihydroxy chalcone derivatives

ABSTRACT

2,4,6-Trihydroxychalcone derivatives of the formula:   WHEREIN R1 is hydrogen, halogen, hydroxy, nitro, alkoxy, amino, alkylamino or dialkylamino and R2 is halogen, jydroxy, alkoxy, nitro, amino, alkylamino or dialkylamino, each of the alkyl portions of the aforesaid alkoxy, alkylamino or dialkylamino radicals having one to three carbon atoms, have hypocholeretic, hypotensive and anti-coagulant properties.

United States Patent Lafon [4 1 Aug. 22, 19712 541 2,4,6-TRIHYDROXYCHALCONE DERIVATIVES [72] Inventor: Louis Lafon, 5 rue de lAlboni,Paris, 16c, France [22] Filed: March 5,1970

[21] Appl.No.: 16,945

[30] Foreign Application Priority Data March 5, I969 Great Britain I1,594/69 [52] US Cl. ..260/590, 260/571, 424/ 331,

' 424/330 [5 l] Int. Cl ..C07c 49/82 [58] Field of Search. ..260/590[56] References Cited FOREIGN PATENTS OR APPLICATIONS 255,188 2 1963Australia ..260/590 Primary Examiner-Daniel D. Horwitz Attorney-Jacobs &Jacobs [57] ABSTRACT 2, 4,6-Trihydroxychalcone derivatives of theformula:

3 Claims, No Drawings 2,4,6-TRIHYDROXY CHALCONE DERIVATIVES Thisinvention relates to 2,4,6-trihydroxy-chalone derivatives, to theirpreparation and to pharmaceutical compositions containing them.-trihydroxy-chalcone The present invention provides, as new compounds,2,4,6-trihydroxy chalcone derivatives of the formula:

OH (I) "-methoxychalcone, 2,4,6-trihydroxy-3',4'-dimethoxychalcone,2,4,6-trihydroxy-3'-nitrochalcone and 2,4,6-trihydroxy-4-nitrochalcone.

The compounds of the present invention have interestingpharmaco-dynarnic activity. More particularly they may be used ashypocholeretics, hypotensives and anticoagulants. Accordingly thepresent invention also provides pharmaceutical compositions comprising a2,4,6-trihydroxychalcone derivative of formula (I) in association with aphysiologically acceptable excipient. For example such compositions maybe in the form of a tablet or capsule containing 0.20 to 0.40 g. of2,4,6- trihydroxy-S'-chlorochalcone or containing'0.20 g. of2,4,6-trihydroxy-3-aminochalcone.

The new compounds can be prepared by condensing phloroacetophenone witha benzaldehyde of the formula:

wherein R, and R are as hereinbefore defined in a solvent mediumespecially acetic acid or ethanol in the presence of hydrochloric acid.The compound obtainedv is generally isolated by precipitation with waterand then it is purified.

The following Examples illustrate the invention.

EXAMPLE 1 2,2,4,4' ,G-PENTAHYDRQXY-CHALCONE OH -Q- l \J In a a stirrer,

16.8 g. (0.1 mol) of dry phloroacetophenone was dissolved in 50 cc. ofslightly warm acetic acid. A solution ofv 13.8 g. (0.1 mol) of resorcylaldehyde in 50 cc. of acetic acid was run into this solution. Thetemperature was raised to between C. and C. Then 4 cc. of dilutehydrochloric acid were added, a complete solution being formed. Stirringwas continued for 1 hour and then the product was precipitated by theaddition of 500 cc. of cold water during which the stirring wascontinued. The precipitate was separated by centrifugation and dried invacuo in the presence of potassium hydroxide. The resulting product 14g.) was purified by treating it with 400 cc. of boiling water to extractany unreacted phloroacetophenone and resorcyl aldehyde. Then the productwas filtered, washed and dried in vacuo in the presence of potassiumhydroxide.

The purified product was collected in the form of a red powder. The dryproduct weighed 9 g. corresponding to a yield of 28 percent. Thisproduct had no welldefined melting point. This compound was soluble insodium hydroxide solution and acetone, slightly soluble in ethyl alcoholand acetic acid and insoluble in diethyl ether, water and benzene.

. EXAMPLE 2 2,3',4,4',6-PENTAHYDROXY-CHALCONE An Erlenmeyer flaskequipped with a stirrer was charged with 50 cc. of acetic acid, 8.4 g.(0.05 mol) of dry phloroacetophenone and 7.5 g. (0.05 mol) of 3,4-dihydroxy-benzaldehyde, and then 3 cc. of concentrated hydrochloric acidwere added. Intense color was formed and then a complete solution wasobtained. This solution was left to stand at room temperature and thenthe-product was precipitated in 500 cc. of cold water. The precipitatewas removed by filtration, washed with water and dried in the presenceof potassium hydroxide.

The dried product weighed 14 g. (a yield of 98 percent) and had no welldefined melting point; This compound was soluble in sodium hydroxidesolution and acetone, slightly soluble in ethyl alcohol and acetic acidand insoluble in diethyl ether, water and benzene.

EXAMPLE '3 2,4,6-TRIl-[YDROXY-3'-AMINO-CHALCONE to stand for 24 hoursand then was poured with stirring into 500 cc. of cold water. Noprecipitation took place. Sodium bicarbonate was gradually added untilpH reached 5. The product was extracted with ethyl acetate and thendried by evaporation.

EXAMPLE 4 2 ,4,6-TRIHYDROXY-4 -DIMETHYLAMINO- CHALCONE OH CH: J CHa-(mil A 200 cc. Erlenmeyer flask equipped with a stirrer was charged with30 cc. of ethyl alcohol, 20 cc. of water, 8.4 g. (0.05 mol) of dryphloroacetophenone and 7.5 g. (0.05 mol) ofpara-dimethylaminobenzaldehyde. 20 cc. of concentrated hydrochloric acidwere then added. This mixture was stirred for 1 hour and then anyunreacted phloroacetophenone was removed by extraction with diethylether. The aqueous layer so obtained was then salted-out by the additionof sodium acetate until the pH reached 5. Extraction with ethyl acetatewas followed by washing in water and evaporation to dryness. The residueso obtained was further purified by the addition of diethyl ether toremove any remaining unreacted compounds. 7.4 g. (i.e., a yield of 35percent) of a compound in the form of a deep violet powder which had nowell defined melting point were obtained. I

This compound was soluble in sodium hydroxide solution, slightly solublein acetone and insoluble in water, hydrochloric acid, diethyl ether,ethyl alcohol, benzene and petroleum ether.

EXAMPLE 5 2,4,6-TRIHYDROXY-3 '-CHLORO-CHALCONE A 200 cc. Erlenmeyerflask was charged with 50 cc. of acetic acid, 8.4 g. (0.05 mol) of dryphloroacetophenone and 7 g. (0.05 mol) of pure 3- chlorobenzaldehyde.This mixture was stirred to form a yellow suspension and then 4 cc. ofconcentrated hydrochloric acid were added to it. A solution formedrapidly, and then after 2 to 3 minutes the solution coagulated. This wasleft to stand overnight and then the product was precipitate in 200 cc.of cold water, removed by filtration, washed in water and dried overpotassium hydroxide.

The dried product weighed 14.2 g., corresponding to a yield of 98percent, and had no well-defined melting point. It was soluble inacetone and sodium hydroxide solution and insoluble in water, .diethylether and benzene.

EXAMPLE 6 2,3,4,6-'I'E'I'RAHYDROXY-4-lvIE'IHOXY-CHAL-- E OH H 0-01)!OOH: l ll A 200 cc. Erlenmeyer flask equipped with a stirrer was chargedwith 50 cc. of acetic acid, 8.4 g. (0.05 mol) of dry phloroacetophenoneand 7.5 g. (0.05 mol) of isovanillin. Partial solution was observed. 3cc. of concentrated hydrochloric acid were added, an intense red colorbeing observed followed by formation of a complete solution within about15 minutes. This solution was left to stand for 24 hours at roomtemperature. Then 500 cc. of cold water were run in resulting in theformation of a salmon-pink precipitate, which was removed by filtration,washed in water and dried in the presence of potassium hydroxide.

The dried product weighed 14.8 g., corresponding to a yield of 98.5percent. Kofler melting point: 250 C. to 260 C. (not very well definedand some decomposition occurred). This product was soluble in sodiumhydroxide solution, acetone, and acetic acid and insoluble in water,diethyl ether and benzene.

EXAMPLE 7 2,4,6-TRIHYDROXY-3 ,4 'DmTl-IOXY-CHAL- CONE p H0@JOCH: I Y Hl.

A 200 cc. Erlenmeyer flask equipped with a stirrer was charged with 50cc. of acetic acid, 8.4 g. (0.05 mol) of dry phloroacetophenone and 8.5g. (0.05 mol) of veratraldehyde. 3 cc. of concentrated hydrochloric acidwere then added. The mixture was left to stand for 24 hours and then wasprecipitated with stirring in 500 cc. of water. The resultingyellow-orange precipitate was removed by filtration, washed and driedover potassium hydroxide.

The dried product weighed 14 g. corresponding to a yield of 88 percent,and had no well defined melting point.

It was soluble in sodium hydroxide solution, acetone and acetic acid andinsoluble in water, diethyl ether, ethyl alcohol and benzene.

EXAMPLE 8 2,4,6-TRIHYDROXY-3 '-NITRO CI-IALCONE A 200 cc. Erlenmeyerflask equipped with a stirrer was charged with 60 cc. of acetic acid,8.4 g. (0.05 mol) of dry phloroacetophenone and 7.55 g. (0.05 mol) ofmeta-nitrobenzaldehyde and stirred for 25 minutes. 3 cc. of concentratedhydrochloric acid were then added and the solution went red. Completesolution was observed and then reprecipitation occurred. The mixture wasleft to stand for 24 hours at room temperature and then 500 cc. of coldwater were run in. The canary-yellow precipitate which formed wasremoved by filtration, washed in water and dried in the presence ofpotassium hydroxide.

The dried product weighed 13.3 g., corresponding to a yield of 88.5percent and its melting point was about 260 C. (not very well definedand some decomposition occurred. It .was soluble in sodium hydroxidesolution and acetone and insoluble in water, diethyl ether, acetic acid,benzene and dimethyl formamide.

I EXAMPLE9 2,4,6-TRIHYDROXY-4'-NITRO CHALCONE A 200 cc. Erlenmeyer flaskequipped with a stirrer wascharged with 60 cc. of acetic acid, 9.35 g.(0.0557 mol) of dry phloroacetophenone and 8.4 g. (0.0557 mol) ofpara-nitro-benzaldehyde. After minutes, solution not being complete, 3cc. of concentrated hydrochloric acid were added. Complete solution andan intense red color were then observed, this being followed byreprecipitation and coagulation. The mixture was left to stand overnightand then poured with stirring into 500 cc. of water. The yellow-orangeprecipitate so formed was removed by filtration, washed in water anddried in vacuo in the presence of potassium hydroxide.

The dried product weighed 14.4 g. corresponding to a yield of 86percent, and its melting point was about 260 C. (not very well defined,and some decomposition occurred). It was soluble in sodium hydroxidesolution and acetone and insoluble in water, diethyl ether, acetic acid,benzene and dimethylformamide.

The pharmacodynamic properties of the new 2,4,6- trihydroxy-chalconederivatives are shown below: 2,2,4,4,6-PENTAHYDROXY CHALCONE When usedin solution form the compound is dissolved in sodium hydroxide solutionand the solution is brought to a pH of 7 by adding hydrochloric acid. 1.Acute toxicity in mice. A Compound used in the form of an aqueoussuspension. When the compound was administered by the gastric route at adosage of 1 g/kg the mortality rate was nil. Sedation and ptosis wereobserved. B Compound in solution form. When the compound wasadministered intraveneously the LI) was 182 i 37 mgjkg.

Symptoms: Diarrhoea, lachrymation, pink coloration for 24 hours in thevein in which the substance has been received. Urine orange colored.Death occurs by pound decreases contractions produced by bariumchloride. Action in conjunction with acetyl choline: At a dosage of 10or 100 y/cc., the compound decreases the contractions produced by acetylcholine.

B In vivo. ileum of guinea pig. Intravenous Injection: When administeredat a dosage of 10 mg./kg. (about one-twentieth of the intravenous LD inmice), the compound arrests peristalsis for 6 to 10 min. The return tonormal is observed to be very rapid.

Note: Marked hypotension, 72 percent on average,

has been observed for 20 to 30 min.

When administered at a dosage of 20 mg./kg. (about one-tenth of theintravenous LI) in mice), the compound arrests peristalsis in an animalfor 20 min., and at the same time 66 percent hypotension is observed for30 min. Inna-duodenal administration: When administered to guinea pigsintraduodenally at a dosage of 100 mg./kg., the compound arrestsperistalsis after 20 to 60 min. The anti-spasm index is percent and atthe same time a hypotension of 25 percent to 40 percent is observed.

3. Action on choleresis in anafistetized rats Intravenous injection ofthe compound at a dosage of 20 mgJkg. (one-tenth of the intravenous LDin mice) produces 33 percent hypocholeresis for 15 min. The bile iscolored red 30 min. after injection.

This compound is particularly efiective as a hypocholeretic.2,3,4,4,6-PENTAI-IYDROXY CHALCONE The compound is used after beingdissolved in sodiurn hydroxide solution and an excess with hydrochloricacid added. 7 1. Acute toxicity in mice When the compound isadministered intravenously the LI) was 82.5 i 3 mgJkg. Sedation andlachrymation are observed.

When it is administered intramuscularly at a dosage of 40 mg./kg.,sedation, hypothermia (2.9), mild q fizips d ld n s sis asi p assassrresl- 2. Cardiovascular properties studied on the isolated heartof a rabbit When the compound was administered at a dosage of ly/ce; thecoronary output increased by from 60 percent to 100 percent. Thiscorresponds to twice the activity of papaverine.

3. Anti-spasmodic action a. Guinea pig ileum in situ: When the compoundwas intravenously administered at a dosage of 8 mg./kg., peristalsis wasreduced, the maximum effect being observed after 30 min. There is 55percent to 70 percent hypotension.

b. Action on Oddis sphincter in dogs: When the compound was administeredintravenously at a dosage of 8 mg./kg., distinct anti-spasmodic actionon the intestine and on Oddis sphincter were observed for 20 min. onaverage.

4. Action on choleresis in anaesthetized dogs Hypocholeretic action wasobserved when the compound was administered intravenously at a dosage of8 mg./kg.

This compound is in particular a hypocholeretic having also ananti-spasmodic action on unstriated muscle fiber. 2,4,6-TRIHYDROXY-3,-AMINO CHALCONE 1. Acute toxicity in the mouse When administered orallyat a dosage of l g./kg. the compound kills two animals out of 12. Deathoccurs 4 days after administration. No symptoms are observed. 2.Tolerance by gastric route in conscious dogs The compound in aqueoussuspension at a concentration of 100 mg./cc. administered orally to dogsat a dosage of 500 mg./kg., in a volume of 5 cc./kg. Tolerance iscomplete and no symptoms are observed. 3. Anti-spasmodic action Guineapig ileum in situ When administered intra-duodenally at a dosage of 125ng/kg. the compound reduces or arrests peristalsis forfrom to more than80 min. The index of peristalsis (number of contractions in 10 min. Xmean amplitude) is reduced on average by 53' percent. Return to normalisobserved. No effect on the arterial bloodpressure is observed.

When administered intra-duodenally at a dosage of 250 mg./kg., thecompound arrests or reduces peristalsis for from to more than 60min.-The index of peristalsis is reduced on average by 88 percent. Ingeneral, the efiect is observed after 9 to min. Return to normal isobserved. v

The compound has an intestinal anti-spasmodic effect on guinea pigs. Italso has an interesting hypocholeretic action, producing 31 percenthypocholeresis in an anaesthetized rat for more than 45 minutes whenadministered intraperitoneally at a dosage of 125 mg./kg.

When taken by man in doses of 0.20 g. four times a day, the compounddisplays a goodeffect on hepatic complaints.2,4,6-TRIHYDROXY4'-DIMETHYLAMINO CHAL- CONE The compound is used in theform of an aqueous suspension.

1. Acute toxicity in mice When the compound is administeredintraperitoneally at the maximum injectable dosage of 400 mg./kg. notoxic effects are observed. The animals display abdominal contractionsand pilo erection. The compound has neither analgesic nor tranquillizingaction.

2. Cardiovascular properties studied on the isolated heart of a rabbit vWhen used in conjunction'with barium chloride, the compound in doses ofl to 100 'y/cc., has the following effects:

a. dilates the coronaries in proportions to the dose;

b. has a negative inotropic effect at 10 and 100 'y/cc.;

c. has a negative chronotropic effect at 10 and 100 'y/cc. 3.Anti-spasmodic properties In vitro: isolated duodenum of rat.

Organs contracted by barium chloride: When administered at a dosage of 1cc. of a 1 percent saturated solution to a 60 cc. cell, the compounddecreases the spasmodic action of the barium.

4. Action on choleresis When intra-peritoneally administered to ananaesthetized rat, in the form of a suspension in water and Tween at adosage of 200 mg./kg., the compound reduces the bile output by 33percent for 15 This compound has particularly hypocholeretic properties.

2,4,6- l ROXY-3'-CHLORO CHALCONE The compound was dissolved in sodiumhydroxide solution (solubility less than 1 percent) and then neutralizedwith hydrochloric acid. it was used as a stable suspension in thepresence of a slight excess of sodium chloride. i

1. Acute toxicity When the compound was administered orally to mice themortality observed was nil at l gJkg. At a dosage of 2 g./kg.administered orally, two animals out of 12 died within 48 hours. Nosymptoms were observed. Observation of animals receiving 600 mg./kg. ofproduct orally: No symptoms were observed, apart from slighthyperthermia of 02.

2. Anti-spasmodic action Guinea pig ileum in situ: When administeredintraduodenally at a dosage of 200 mg./kg., the compound arrests orreduces peristalsis. The index of peristalsis drops on average by 70percent for from 20 to more interesting than min. At the same time, l2percent to 62 percent hypertension was found in 3 experiments and 25percent hypotension in the fourth experiment. Anti-spasmodic action indogs: When administered intra-duodenally at a dosage of 200 mg./ kg., to4 dogs subjected to the action of prostigmine morphine, the compound hadan anti-spasmodic action. There was a reduction in the intestinalperistalsis with a slight increase in tone, for about 20 min. Variationsin the forcing pressure of Oddis sphincter were reduced on average by 40percent for a period varying between 30 and 60 min. 3. Action oncholeresis The compound was administered intra-duodenally at a dosage of200 mg./kg., to 2 sets of anaesthetized rats. In the first set, 43percent hypocholeresis was observed for 15 mins.; and in the second set,29 percent hypocholeresis for 15 min. 7 r

The compound has also been tested, as regards its action on choleresis,on dogs, at a dosage of 200 mg./kg.,

of 38 percent in the bile output was observed. This reduction lastedmore than half an hour.

This compound is particularly effective as a hypocholeretic.

For use in the treatment of man, the compound may be used in the form ofcapsules or tablets containing 0.20 to 0.40 g. of the active ingredientto be ad ministered 3 to 6 times per day. Clinical tests confirm thepharmacological tests performed on animals.

The compound was preventatively administered at a dose of 3 capsules,each containing 0.20 g. of the compound, to 10 patients who had beensubjected to a morphine-choleretic test (Test of du Fablet, Debray andHardouin). In 8 cases the occurrence of biliary pain and bilious attackswas suppressed.

' Used at a dosage of 3 capsules per day the compound gives excellentresults in the treatment of biliary pain and bilious attacks.

2,3 ,4 ,6-TETRAHYDROXY-4'- -METHOXY CHAL- CONE The compound is dissolvedin sodium hydroxide solution, the excess being neutralized withhydrochloric acid (final pH 7).

l Acute toxicity in mice When administered intravenously the compound hObservation on animals receiving 35 mg./kg. of the compoundintramuscularly.

Sedation, dypsnoea, haematuria, hypothermia (2.8 and mydriasis areobserved. The compound has some slight analgesic action and a moderatetranquillizing effeet.

2. Cardiovascular properties studied on the isolated heart of a rabbitThe compound was used in solution in the Van Dyke Hasting liquidperfusing the heart.

In hearts constricted with barium chloride, the compound, when tested atdosages of 100 and 10 y/cc. dilates the coronan'es, has a slightpositive inotropic action, does not alter the rate of the heart beat at10 y/cc., but reduces it (50 percent) at 100 'y/cc.

As regards its coronary dilation, the product is twice as active aspapaverine (88 percent and 74 percent dilation of 2 'y/cc.). (Toxicityof papaverine when administered intravenously: LD 36 mg./kg. Thiscompound has useful coronary dilating properties.

3. Experiments on dogs A chloralosed animal was given intravenously adose of mg./kg. l-lypotension was observed, lasting on average 45 min.,at a level of from 20 percent to 30 percent of the dogs initial arterialpressure. 2,4,6-TRlHYDROXY-3,4-DIMETHOXY CHAL- CONE l Acute toxicity inmice When administered intravenously the compound has an LD of 97 i 8mg./kg.

Symptoms: Exophthalmia, diarrhoea, bleeding at site of injection for anextended time.

With very high dosages (340 mg./kg.), nasal and oral hemorrhages wereobserved. Autopsy showed the lungs apparently hemorrhaged.

Observation on animals receiving 52.5 mgJkg. of the compoundintramuscularly No symptoms apart from slight hypothermia (0.7) wereobserved.

2. Anti-spasmodic properties A In vitro isolated I duodenum of rat.Organs in relaxed state When administered at a dosage of l mg./cc., thecompound reduces the organ contraction by 20 percent to 30 percent.

B In vivo Guinea pig ileum in situ.

When injected intravenously into guinea pigs at a dosage of 10 mg./kg.(one-tenth the intravenous LD the compound arrests peristalsis for 2 to15 min., this being followed by frequency stimulation.

Marked hypotension, amounting to about 60 percent, is also observed.2,4.6-TRIHYDROXY-3 '-NITRO CHADCONE The compound is dissolved in sodiumhydroxide solution and the excess neutralized with hydrochloric acid.

When administered intravenously to mice the compound has an LD of 71 i 5mg./kg. The animals die from acute oedema of the lung.

When administered at a dosage of 1 mg. /kg., the compound produces verymarked hypotension 'in guinea pigs in vivo.

When administered intravenously to dogs at a dosage of 5 mg./kg. thecompound produces 20 percent to 40 percent hypotension, which lasts onaverage one hour. 2,4,6-TRII-IYDROXY-4'-NH'I'RO ONE The compound isdissolved in sodium hydroxide solution and neutralized with hydrochloricacid.

1. Acute toxicity in mice When the compound is administeredintravenously at a dosage of mg./kg., a mortality rate of 3/12 of theanimals is observed.

Symptoms: sedation, pilo-erection, lachrymation and diarrhoea.

When the compound in crude form (i.e., not in solution) is administeredorally to mice at a dosage of 400 mg./kg. it produces nil mortality. 2.Anti-coagulant action A study of anti-coagulant action was carried outon rabbits. The compound has a distinct anti-vitamin-K action which isobserved after the ingestion of 2 g./kg. of the product.

This compound has advantages as an anti-coagulant. In the treatment ofman, it may be used in the form of a capsule or tablet containing 0.05g. to 0.10 g., the dose being from 3 to 4 capsules or tablets per day.

I claim:

1. 2,4,6-Trihydroxy-chalcone derivatives of the formula:

wherein R, is hydrogen and R is nitro.

2. A 2,4,6-trihydroxychalcone derivative according to claim 1 which is2,4,6-trihydroxy-3-nitrochalcone.

3. A 2,4,6-trihydroxychalcone derivative according to claim I which is2,4,6-trihydroxy-4'-nitrochalcone.

2. A 2,4,6-trihydroxychalcone derivative according to claim 1 which is2,4,6-trihydroxy-3''-nitrochalcone.
 3. A 2,4,6-trihydroxychalconederivative according to claim 1 which is2,4,6-trihydroxy-4''-nitrochalcone.